This invention relates to the isolation of two compounds namely (xe2x88x92)- Matairesinol (4,41-dihydroxy-3, 31-dimethoxylignan-9, 91-olide) and (xe2x88x92) wikstromol (4,41,8-trihydroxy-3, 31-dimethoxyliganan-9, 91-olide) from the plant source Cedrus deodara in significant yields. This invention also identifies the use of the said compounds as an antioxidant or a free radical scavenger.
The compound (xe2x88x92)-Matairesinol and (xe2x88x92)-wikstromol as such do not find much commercial value. The structural formulae of (xe2x88x92)-Matairesinol and (xe2x88x92)-wikstromol are as shown in FIG. 1. Lignans are widely distributed in angiosperms and gymnosperms. The range of their structures and biological activities is broad. Literature indicates that Matairesinol exhibits CAMP phosphodiesterase inhibition activity, [W. Donald Macrae and G. H. Neil Towers, Biological activities of Lignans, Phytochemistry 23 (6), 1207-20 (1984)]. (xe2x88x92)-Wikstromol is found to be active against p-388 lymphocyte leukemia and anti-HIV activity (M. K. Kharmlach, R. Dhal and E. Brown, Premieres syntheses totales du (+)-wikstromol, de la (xe2x88x92)-Trachelogenine, De la (xe2x88x92)-Nortrachelogenine et des lignoides apparentesxe2x80x94Tetrahedran 48, 10115-10126 (1992)).
The wood of Cedrus deodar possesses diaphoretic, diuretic and carminative properties, and it is useful in treatment of fevers, piles and pulmonary and urinary disorders. The extract of the bark is astringent and useful for fevers, diarrhea and dysentery. The oleoresin of deodar and the dark-colored oil obtained from the wood are valued for their application for ulcers and skin diseases. [Ref: Wealth of India, Vol.II, P.108-10 (1950) (published by CSIR)].
A close look at literature suggests that the lignan, seco-isolariciresinalldiglycoside (SDG) which has been reported to possess multitude of activities is isolated from [U.S. Pat. No. 5,846,944]. The natural lignans (xe2x88x92)-matairesinol reduce the binding of H-labeled 5xcex1-dihydrotestosterone (DHT) to human sex hormone-binding globulin (SHBG). [Matthias Schottnerand Gerhard Spiteller (J. Nat. Prod. 1998, 61, 119-121)]. P. K. Agarwal and R. P. Rastogi (Phytochemistry Vol 21, No 6, pp 1459-1461, 1982) reported isolation of two lignans meso-secoisolariciresinol and cedrusinin from Cedrus deodara. 
The isolation of matariesional in 0.10% yield and (+)-wikstromol in 0.124% yield is reported from Wikstroemia viridiflora (Wikstromol, a new lignan from Wikstroemia viridifloraxe2x80x94Sheela Tandon and R. P. Rastogi; Phytochemistry 1976, vol 15, pp 1789-1791).
The extraction of the lignans and other constituents namely (xe2x88x92)-nortrachelogenin, carinol and carissanol from Carissa edulis has been reported by Hans Achenbach, Reiner Waibel and Ivan Addae-Mensah, Ref. Phytochemistry, Vol. 22, No. 3, pp. 749-753, 1983.
There is a considerable amount of epidemiological evidence indicating an association between diet rich in fruits and vegetables and a decreased risk of cardiovascular disease and certain forms of cancer. It is generally assumed that the active principles contributing to these protective effects are nothing but antioxidant phytochemicals.
Recent research is directed to find out phytochemicals from plant sources in highlighting the role of polyphenolic compounds of plant materials as antioxidants, antimutagenic, anti inflammatory, antiatherosclerotic, antidiabetic, antihepatotoxic and antimicrobial agents. [Overview of flax lignans by Neil D. Westcott and Alister D. Muir, Crop Utilization Section, Saskatoon Research Centre, Agriculture and Agri-Food Canada, 107, Science Place, Saskatoon, SK S7N 0X2, Canada in Volume IIxe2x80x94January 2000xe2x80x94inform]. Alcoholic extract of stem C. deodara was found to have anti-cancer activity. [Ref. Medicinal Plants of India (ICMR) Vol.I, 1976, pp. 214 and 215].
Medicinal importance of antioxidant active principles are acquiring importance because of the involvement of free radical mediated oxidative stress in age related diseases.
Free radicals are highly energized molecules that contain an unpaired electron. These are produced through normal biological and environmental processes involving oxygen and can trigger chain reactions that product more free radicals. Normally there is a balance between the amount of free radicals generated in the body and antioxidants to protect against them. However, natural antioxidants present in the body can cope up only with the optimal generation of free radicals and any additional burden of free radicals or lack of antioxidant protection can tip this balance and lead to oxidative stress.
There is a need for free radical scavengers or antioxidant principles isolated from natural resources for preparation of formulations useful in alleviating diseases. Hence, it becomes necessary to look for other lignans possessing related and important biological properties.
Accordingly, the applicants conducted a detailed study on principles from Cedrus deodara and this investigation led to the isolation of two active principles namely, Matairesinol and wikstromol. These compounds although hitherto isolated from Wikstromia spp. were in low yields. Cedrus deodara hence is a new source for these lignans and their presence in this taxon in significantly high yields makes this invention more important.
The main object of the invention relates to use of (xe2x88x92)-wikstromol and/or (xe2x88x92)-matairesinol as free radical scavengers or antioxidants:
Another object of the invention is to provide methods for the isolation of (xe2x88x92)-wikstromol and/or (xe2x88x92)-matairesinol from Cedrus deodara. 
Still another object of the invention is to provide methods for the treatment of arteriosclerosis.
Accordingly, the invention provides novel compositions containing (xe2x88x92)-wikstromol and/or (xe2x88x92)-matairesinol and useful as free radical scavengers and antioxidants. The invention further provides methods for the isolation of (xe2x88x92)-wikstromol and/or (xe2x88x92)-matairesinol from Cedrus deodara as well as use of the compounds for the treatment of arteriosclerosis.
Accordingly, the invention provides a composition comprising an effective amount of (xe2x88x92)-wikstromol together with or associated with an additive and useful as an antioxidant.
In yet another embodiment the composition contains an effective amount of (xe2x88x92)-matairesinol together with additives and useful as an antioxidant.
In another embodiment the additive is selected in such a manner that they has no effect with the active principles of the composition such as nutrients etc., The additive is such that they enhance and do not retard the activity of the active ingredients i.e. (xe2x88x92)-wikstromol and/or (xe2x88x92)-matairesinol.
In another embodiment the additive is selected from nutrients such as carbohydrates, proteins, sugar and pharmaceutically acceptable carriers.
In another embodiment the ratio of (xe2x88x92)-wikstromol and/or (xe2x88x92)-matairesinol with the additive is in the range between 0.1:10 to 2:10.
In an embodiment (xe2x88x92)-wikstromol and/or (xe2x88x92)-Matairesinol present in an amount of 250-300 mg.
Further the invention provides a process for the isolation of (xe2x88x92)-wikstromol and/or (xe2x88x92)-matairesinol from the Cedrus deodara, said process comprising the steps of
a) extraction of the pulverized plant parts of Cedrus deodara with solvents to remove the essential oils;
b) concentrating the extract under vacuum to obtain a residue;
c) adding ethyl acetate to the residue obtained in step (b);
d) separating the solvents by conventional methods;
e) subjecting the residue to a first elution with about 3% methanol in chloroform to obtain (xe2x88x92)-matairesinol; and
f) subjecting the residue of step (e) to a second elution with about 5% methanol to obtain (xe2x88x92)-wikstromol.
The solvents used in step (a) are hexane and chloroform.
In another embodiment, the plant parts of Cedrus deodara such as bark and leaves are used for extraction.
In yet another embodiment, the wasted plant parts of Cedrus deodara are employed for isolation of the said compound. Preferably, the waste left after extraction of essential oil from the plant parts is used in the process.
There is enough literature, for example the article in Inform Vol. 11, pg. 118-121 that suggests that oxygen free radicals are responsible for development of arteriosclerosis. And, that, antioxidants are useful to retard hypercholestrolemic arteriosclerosis.
Accordingly, it is a further feature of the invention to treat arteriosclerosis wherein an effective amount of (xe2x88x92)-wikstromol or (xe2x88x92)-matairesinol is administered to a subject in need thereof.
(xe2x88x92)-wikstromol or (xe2x88x92)-matairesinol may be administered together with or in combination with therapeutically acceptable additives. The effective amount of (xe2x88x92)-wikstromol or (xe2x88x92)-matairesinol that may be administered to a subject can be readily determined by a person skilled in the art. However, it is recommended that the dosage of (xe2x88x92)-wikstromol or (xe2x88x92)-matairesinol administered may be in the range of 250 to 300 mg per dose, twice a day. While it is possible to administer the composition in oral as well as systemic routes, the oral route achieves the desired best results.
Compositions employing (xe2x88x92)-wikstromol or (xe2x88x92)-matairesinol may be prepared by conventional methods as known in the art. The compositions may be in the form of tablets, capsules or syrups, etc. Suitable additives as known in the art may be selected for the preparation of these compositions.
In essence the focus of the invention is to provide methods for using (xe2x88x92)-wikstromol and/or (xe2x88x92)-matairesinol for the preparation of compositions useful as a free radical scavenger and antioxidant.
The heartwood of Cedrus deodara finds extensive use in essential oil industry. The oil by name xe2x80x98cedar woodxe2x80x99 oil finds application in flavor and fragrances. The heartwood powder after extraction of essential oil is a by-product and waste. This invention relates to isolation and purification of the compounds (xe2x88x92)-matairesinol and (xe2x88x92)-wikstromol and use as antioxidants and for the treatment of arteriosclerosis.
This present invention relates to the isolation of two compounds namely (xe2x88x92)-Matairesinol and (4,41-dihydroxy-3, 31-dimethoxylignan-9, 91-olide) and (xe2x88x92)-wikstromol (4,41,8-trihydroxy-3, 31-dimethoxyliganan-9, 91-olide) from a new source Cedrus deodara. This invention also relates to new use of the compounds as an antioxidant.
The present invention embodies isolation of (xe2x88x92)-Matairesinol and (xe2x88x92)-wikstromol, two antioxidant principles from an entirely new source and their free radical scavenging property compared with known biologically proved anti free radical agents, anti dyslipidemic and anti hepatotoxic efficacies.